Accepted Manuscript
Chemical composition antibacterial and antifungal activities of flowerhead and root essential oils of Santolina chamaecyparissus L., growing wild in Tunisia
Karima Bel Hadj Salah-Fatnassi, Faten Hassayoun, Imed Cheraif, Saba Khan, Hichem Ben Jannet, Mohamed Hammami, Mahjoub Aouni, Fethia Harzallah- Skhiri
PII: S1319-562X(16)30001-8 DOI: http://dx.doi.org/10.1016/j.sjbs.2016.03.005 Reference: SJBS 692
To appear in: Saudi Journal of Biological Sciences
Received Date: 27 February 2016 Revised Date: 21 March 2016 Accepted Date: 22 March 2016
Please cite this article as: K.B.H. Salah-Fatnassi, F. Hassayoun, I. Cheraif, S. Khan, H.B. Jannet, M. Hammami, M. Aouni, F. Harzallah-Skhiri, Chemical composition antibacterial and antifungal activities of flowerhead and root essential oils of Santolina chamaecyparissus L., growing wild in Tunisia, Saudi Journal of Biological Sciences (2016), doi: http://dx.doi.org/10.1016/j.sjbs.2016.03.005
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root essential oils of Santolina chamaecyparissus L., growing wild in Tunisia
Karima Bel Hadj Salah-Fatnassi a,b*, Faten Hassayoun a, Imed Cheraif c, Saba Khanb,
Hichem Ben Jannet d, Mohamed Hammami c, Mahjoub Aouni a, Fethia Harzallah-
Skhiri e a Laboratoire des maladies transmissibles et substances biologiquement actives LR99-ES27, Faculté de Pharmacie Monastir, Université de Monastir, Tunisie b Biological Sciences Department, Faculty of Science & Art -Rabigh, King Abdulaziz University
(KAU), Saudi Arabia c Laboratoire de Biochimie, USCR de Spectrométrie de Masse, Faculté de Médecine, Université de
Monastir, Tunisie d Laboratoire de chimie des substances naturelles et de synthèse organique, faculté des sciences,
Université de Monastir, Tunisie e Laboratoire de Génétique, Biodiversité et Valorisation des Bioressources, Institut Supérieur de
Biotechnologie, Université de Monastir, Tunisie.
* Corresponding author
Dr. Karima BEL HADJ SALAH Biological Sciences Department Faculty of Science & Art- Rabigh 21911 King Abdulaziz University. Saudi Arabia Tel.; +966 0507634489 E-mail address; [email protected]
1. Introduction
A growing body of evidences suggested that more than three antibiotics derived from microorganisms are initiated every year (Clark, 1996, Rudi et al., 2012). Novel resources, especially plant, are also being examined. Essential oils, secondary metabolites and medicinally important compounds with or without bioactivity, have been isolated from plants belonging to the Asteraceae family: Achillea L., Anthemis L., Artemisia L., Balsamita Desf.,
Chrysanthemum L., Matricaria L., Tanacetum L., and Santolina Tourn (Khallouki et al.,
2000). Besides the different strains, several genuses from the family Asteraceae, have been particularly shown to be antimicrobial (Paulo, 2006; Salie et al. 1996; Wetungu et al.,
2014).
In Mediterranean area, it has been found that the genus Santolina (Asteraceae, tribe
Anthemideae) is characterized more than 10 species that are distributed widely (Derbesy et al., 1998). Usually plants of the genus Santolina cultivate in South Europe and North Africa.
As a result, numerous class of this taxon have been investigated biologically and chemically and give rise to number of mono and sesquiterpenoids along with some other secondary metabolites (Barrero et al., 1998; Barrero et al., 1999; Barrero et al., 2000; Marco et al. 1993).
In all these Santolina species, S. viridis Wild (South of France and North of Spain), S. pectinata Lag. (Iberian peninsula) and S. chamaecyparissus L. (Asteraceae) are the most widely spread species around the world. S. chamaecyparissus L. is synonyme of Ormenis africana (Jord. et Fourr.) Out of four species of the genus Ormenis only O. africana is vivacious (Poli et al., 1997) and others are used in important medicine (Pons Giner & Rios
Canavate., 2000). Widely known shrub (S. Chamaecyparissus) with yellow inflorescences utilized in Mediterranean folk medicine because of its analgesic, bactericidal, fungicidal, vermifuge and vulnerary properties (Cuéllar et al., 1998; Grieve, 1984; Bean, 1989). Also it has been suggested that this plant is utilized in phytotherapy for numerous kinds of dermatitis
(Giner et al. 1988) and also as a stimulant and a stomachic (Yoganarasimhan, 2000).
Although preliminary evidence available in the literature suggested that S. chamaecyparissus has anti-inflammatory activity and anti-phospholipase A2 due to the presence of the isolated active principle nepetin present in the dichloromethane extract (Sala et al., 2000). In Tunisia,
S. chamaecyparissus is the only Santolina species recorded and is used as vermifuge and emmanagogue (Le Floc’h, 1983).
The components of essential oils from diverse species of Santolina have been investigated: S. oblongifolia (De Pascual et al., 1983), S. ligustica (Flamini et al., 1999), S. rosmarinifolia
(Pala-Paul et al., 2001), S. canescens (Casado et al., 2001) and S. chamaecyparissus (Garg et al., 2001; Lawrence, 1997) , and all of these species produced monoterpene oils and demonstrated diverse constituents.
Several volatile oils are recognized to acquire antifungal and antibacterial properties and are potentially applicable as antimycotic agents. Many scientific investigations have been conducted to evaluate the biological activities of about 60% of the essential oils possess antifungal and 35% of them exhibited antibacterial properties (Chaurasia & Vyas, 1977). A few series of studies has demonstrated the potential antimicrobial effect of essential oils from various Santolina species, i.e. Santolina corsica oil exhibited an appreciable antibacterial activity against Staphylococcus aureus (Rossi and al., 2007) sesquiterpenes from Santolina rosmarinifolia showed significant antimicrobial activity against yeasts while none of the compounds tested showed significant activity besides Gram positive and Gram negative bacteria (Barrero et al., 1999).
To specifically address the constituents of an essential oil and antimicrobial properties of
Tunisian S. Chamaecyparissus, we directly examined to investigate the chemical composition, antibacterial and antifungal activities of essential oils isolated by hydrodistillation from the flowerheads and roots of S. chamaecyparissus L., growing wildly in Tunisia.
2. Methodology
2.1 Plant material Plants of S. chamaecyparissus were collected at the time of flowering stage from Siliana on the mountain ‘Djebel Kesra’ located in the West Center of Tunisia at 35.81 (latitude in decimal degrees), 9.36444 (longitude in decimal degrees). The identification of plant material has been demonstrated according to the flora of Tunisia (Pottier-Alapetite, 1981) by the botanist Dr. Fethia Harzallah-Skhiri (Higher Institute of biotechnology, Monastir University,
Tunisia)
2.2 Plant extraction and essential oil analysis a) Plant extraction
Essential oils from the flower heads and roots of S. chamaecyparissus were found by hydrodistillation for 4 h by using a Clevenger type apparatus (Benyelles & al. 2014; Suwei & al. 2014; Okoh & al. 2010). Anhydrous sodium sulphate served to dry essential oils and stored at 4 °C until use. The yield was calculated according to the plant fresh weight. b) Essential oil analysis
The identification of essential oil components was performed by GC and GC/MS. GC diagnostic were carried out on an HP5890-series II gas chromatograph (Agilent Technology,
California, USA) with Flame Ionization Detectors (FID) characterised by: a fused silica capillary column, non polar HP-5 and polar HP Innowax (30 m x 0.25 mm ID, film thickness
0.25 µm). The temperature of the oven was 50 °C for 1 min, then it is programmed to achieve heating rate up to 5 to 240 °C/min and held isothermal for 4 min. Injector temperature: 250
°C, detector temperature: 280 °C; nitrogen used as a carrier gas (1.2 ml/min); and injected 0.1 ml diluted in hexane1%. The percentage of the constituents was calculated by electronic integration of FID peak areas without the use of response factor correction. Hewlett-Packard
5972 MSD system served for GC/MS analyses: HP-5 MS capillary column (30 m x0.25 mm
ID, film thickness of 0.25 mm) was directly coupled to the mass spectrometry. The comparison of the components mass spectra with those in the Wiley 275 GC/MS library and of their retention indices with literature data (Adams, 2001) served to their identification.
Retention indices (RI) were calculated by the retention times of a series of n-alkanes.
2.3 Antibacterial activity a) Bacterial strains
The antibacterial assays were carried against four gram negative bacteria: Esherichia coli
ATCC 25922, Pseudomenas aeruginosa ATCC 27853, Proteus mirabilis and Citobacter freundeii and two gram positive bacteria: Enterococcus faecalis ATCC 29212 and
Staphylococcus aureus ATCC 25923 (American Type Culture Collection, Rockville, MD).
Inoculums were kept in nutrient agar (Sigma) at 37°C. Bacterial suspension were prepared in
Mueller Hinton Broth (Sigma) and adjusted to a108 cfu/ml for overnight incubation,
b) Agar diffusion assay
Antibacterial studies have been evaluated by the method of disc-diffusion (Sahin et al., 2003).
One millilitre from a bacterial suspension of 108 cfu/ml was spread on the surface of both control and test plates.
Under aseptic conditions, 10 µl of essential oil were applied to sterilized Whatman filter paper discs N° 3 (6 mm diameter) and placed on the agar surface. Before incubated at 37 ºC, plates were left for 2-3 hours at 4 °C to help the diffusion of the oil. Diameters of inhibition were measured in millimetre. Gentamicin (10µg/disque; Gibco) was used as standard antibiotic for comparison.
C )Micro-well dilution assay
The flowerhead and root oils from S. chamaecyparissus were diluted in ethanol 99° (v/v) to the highest tested concentration of 10 µg/ml. Serial two-fold dilutions (0.078 to 10 µg/ml) of oils were prepared using nutrient broth. Bacterial strains were inoculated for 12 h broth cultures and suspensions were adjusted to 0.5 McFarland standard turbidity. The Minimal
Inhibition Concentration (MIC) defined as the lowest concentration of the essential oil to inhibit the growth of microorganisms and was determined on the basis of micro-well dilution method (Gulluce et al., 2004a, b,).
2.4 Antifungal testing a) Fungal strains
Seven strains of fungi were used for the antifungal tests, comprising: three dermatophytes
(Trichophyton rubrum, Microsporum canis and Epidermophyton floccosum); one opportunist pathogenic yeasts (Candida albicans); and three hyphamycets (Scytalidium dimidiatum,
Scopulariopsis brevicaulis and Aspergillus fumigatus). The micro-organisms were collected from Pasteur Institut, Paris (France) or the Microbiology Laboratory, Faculty of Medecine,
Besançon (France).
a) Agar incorporation method
The antifungal assays of Santolina essential oils were carried out by agar incorporation method (Benjilali et al., 1986; Yang et al., 1996; Griffin et al., 2000). The essential oils were dissolved in 99% EtOH this solvent was used as a negative control. Then a precise volume is mixed aseptically with 100 ml of Sabouraud glucose agar (SGA) to give final concentrations of 1000, 750, 500, and 250 µLml-1. Dermatophytes were inoculated by disposing 5 mm mycelium in the center of plates while C. albicans by adding 1ml from a blastospore suspension. The Petri dishes were then incubated for 24 h at 37°C for Candida and
Aspergillus and at 24°C for 7 days for dermatophytes and Scopulariopsis. These tests were carried out in triplicates. Two methods served in the evaluation of the antifungal activity of the essential oil: 1- the minimal inhibitory concentration (MIC), the lowest concentration which inhibits the visible growth of fungi during the incubation period.
2- the percentage inhibition 1% according to the method of Singh et al. (1993).
3. Results
Chemical composition of the essential oil
With the help of hydrodistillation method of flowerheads, picked from S. chamaecyparissus had yellow colour and an agreeable smell, the yield oil was 0.062% (v/w), volume/fresh weight), as a result of which essential oils is obtained. Whereas the root oil had orange-brown colour and a pungent odour with 0.148% yield oil. The results obtained by qualitative and quantitative analysis by GC and GC/MS are shown in Table 1.
All the compounds acquired are listed in order of their elution. Sixty seven constituents were identified representing 99.14% and 99.44% of the total S. Chamaecyparissus flowerhead and root oil respectively. The prevalent constituents were 1,8-cineole (12.94%) β-eudesmol
(10.49%), terpinene-4-ol (6.97%), γ-cadinene (6.55%), spathulenol (5.80%), camphor
(5.27%), germacrene-D (5.03%) and myrtenol (4.26%). Oxygenated monoterpenes represented 36.94% of the total oil, 1.8 cineole (12.9%) and terpinen-4-ol (6.9%) the most abundant compound. Significant amounts of oxygenated sesquiterpenes (29.8%) mainly represented by β-Eudesmol (10.9%) and spathulenol (5.8%). Sesquiterpene hydrocarbons
(16.8%) were also observed. The monoterpene hydrocarbons represented only 7.2% of the total oil.
Antifungal and antibacterial activity
Antifungal activity of the flowerhead and the root essential oils has been reported in Table 2.
In the presence of 500 µg/ml of essential oil, the inhibition rate varied from 0 to 89.25%. Out of three dermatophytes tested, the flowerhead oil showed a very strong inhibition rate (73 -
89.25%); whereas root essential oil showed an average to strong inhibition rate (29.03 -
68.00%). Epidermophyton floccosum which was the most resistant to the root essential oil
(29.03%) was the most sensible to the flowerhead oil (89.25 %). We have noticed that there is no inhibition on the pathogenic yeast Candida albicans at 500 µg/ml, though it was sensible at 1000 µg/ml. The MIC varied from 500 to 1000 µg/ml and from 750 to1000 µg/ml respectively for the flowerhead and the root essential oils of S. chamaecyparissus (Table 2).
According to the results of the agar diffusion method, Enterococcus faecalis (ATCC 29212) was the most susceptible microorganism which was strongly inhibited by the flowerhead oil (inhibition zone was 26 mm) (Table 3). As it has been shown in the antifungal tests, bacteria seem to be more sensible to flowerhead than root essential oil.
The concentration rate of MIC has been reported in Table 3. Both EOs of S. chamaecyparissus delineate antibacterial activity against all bacterial strains, with MIC values ranging from 0.625 to10 µg/ml. None of the essential oil tested inhibited Pseudomonas aeruginosa ATCC 27853 by the disc diffusion method, though this germ was sensible to both flowerhead and root Eos by microwell dilution method with MIC of 0.626 µg/ml and 5 µg/ml respectively.
4. Discussion
Compared with previous reports, the composition of the oil that we analyzed greatly differed from the oils of S. chamaecyparissus L. collected in Egypt (Aboutabl, et al., 1987), in France
(Vernin, 1991), in Italy (Tognolini et al, 2006) and in Algeria (Nouasri et al., 2015)(44.5%,
45%, 28.24% and 40.33% respectively). Valencia and Djeddi et al., 2012 studied that
Santolina chamaecyparissus L. of Algeria showed a different distribution of components with camphor as the main component at 25% and 31.1% respectively.
In previous reports the oils of S. canescens, S. rosmarinifolia and S. oblongifolia were characterized by santolindiacetylene, acetylenic, and dihydrofuran sesquiterpenes respectively with some interesting similarities existing between the oils of Tunisian S. chamaecyparissus and S. ligustica, which oil was characterized by myrcene, 1,8-cineole, and terpinen-4-ol
(Tirillini et al., 2007) . Moreover, key compounds analysed in the oil from flowerheads of S. chamaecyparissus differ from those detected in Tunisian essential oil (Terpinene-4-ol (34%),
Borneol (17%), Germacrene D (5%) and γ-Terpinene (7%)) (Fridlender et al., 2002).
In our study, S. chamaecyparissus essential oil demonstrated variability in the quantitative and qualitative contents with the same species growing in other geographic areas. The composition of plants analysed in the present study was close to that of S. chamaecyparissus
L. sp. squarrosa from Spain (p-cymene, 5%), camphor, 25.19%), (bornyl acetate, 9.9%), allo- aromadendrene (19.04%), α-muurolene (7.28%), artemisia cetone (0.64%)) (Villar et al.,
1986).
Further investigations of the essential oil composition for a larger number of populations of this species, along with more data on the different species of the genus Santolina, can be helpful in chemiotaxonomy.
Eo from Santolina seems to be more active when using MIC method than disc diffusion method. Agar diffusion technique is considered as less suitable to estimate the antimicrobial activity of EOs since the active volatile components are likely to be evaporated together with the dispersing solvent, and their non polar nature prevents them from diffusion through the agar media (Goñi et al., 2009).
As per our knowledge, the antibacterial and antifungal actions of S. chamaecyparissus flowerhead and root essential oils, has been reported for the first time. However, few studies on antimicrobial activities of S. chamaecyparissus were previously described. An essential oil obtained from the herb of S. chamaecyparissus L. is effective in controlling candidiasis both in vivo (Suresh et al., 1995), and in vitro (Djeddi et al., 2012; Nouasri et al., 2015); while the dichloromethane extracts of S. chamaecyparissus L. subsp. squarrosa exhibited interesting inhibition against Rhizopus stolonifer (Lopez et al., 2008).
Several components detected in the Tunisian S. chamaecyparissus flowerheads essential oil have been reported as efficient antibacterial or antifungal agents, such as 1,8-cineole
(Cimanga et al., 2002; Tzakou et al., 2001; Pattnaik et al., 1997; Tirillini et al., 1996), α- terpineol, terpinen-4-ol, α-pinene, β-pinene, α-phellandrene, p-cymene (Dorman & Deans,
2000). It has been demonstrated in the literature that the inhibitory activity of an essential oil results from a complex interaction between its different constituents, which may produce additive, synergistic or antagonistic effects, even for those present at low concentrations
Xianfei et al., 2007; Zakarya et al., 1993), i.e. 1,8-Cineole in combination with camphor has shown higher antimicrobial effects (Viljoen et al., 2003) . On the other hand, the compounds present in the greatest proportions are not necessarily responsible for the greatest share of the total activity, and then, the activity could be attributed to the presence of minor components such borneol (3.66%), carvacrol (2.51%) and myrtenal (0.66%) known already to exhibit an antibacterial activity (Zakarya et al., 1993; Viljoen et al., 2003; Onawunmi, 1984).
The type of action of antimicrobial agents depends on the kind of microorganism and evidence designated that in the case of essential oils, it is largely associated with cell membrane damage. Their chemical components are characteristically hydrophobic and will accumulate in the lipid-rich environments of cell membrane structures and cause structural and functional damage. On the other hand, hydrophobicity and ability to damage cell membrane structures are not the only factors involved and it is obvious that toxicity is linked to an optimum range of hydrophobicity (Cox et al., 2000; Lambert et al., 2001).
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Table1. Composition of S. chamaecyparissus flower head and root essential oils.
N° Compounds RI a Percentage
Flowerhead Root
1 2-Butanone 896 0.04 -
2 α-pinene 1020 0.09 0.02
3 Camphene 1071 0.09 0.02
4 β-Pinene 1113 0.88 0.04
5 Sabinene 1124 0.56 -
6 Myrcene 1161 2.96 0.10
7 α-Phellandrene 1168 0.22 -
8 α-Terpinene 1186 0.22 -
9 Limonene 1196 0.15 -
10 1,8-Cineole 1214 12.94 0.03
11 (z)-β-Ocimene 1235 0.00 -
12 δ-Terpinene 1245 1.04 0.03
13 p-Cymene 1269 0.79 -
14 Terpinolene 1286 0.16 -
15 Isoamyl isovalerate 1287 0.29 -
16 Perillene 1306 0.18 -
17 E-2-hexenyl acetate 1327 - 0.03
18 Yomogi Alcohol 1382 0.03 -
19 Fenchone 1404 0.03 -
20 Artemisia cetone 1410 0.13 0.06
21 Camphene hydrate 1442 0.06 -
22 Ylangene 1452 0.17 -
23 α-Cubebene 1459 0.05 - 24 α-Ylangene 1467 0.07 -
25 Camphanolene aldehyde 1470 0.09 -
26 α-Copaene 1489 0.01 0.83
27 Artemisia alcohol 1495 0.08 0.03
28 α-Gurjunene 1510 0.08 0.06
29 Camphor 1517 5.27 0.10
30 Linalool 1548 0.33 -
31 Linalyl acetate 1554 0.47 -
32 Elemene 1555 0.13 -
33 Isobornyl acetate 1582 0.50 0.04
34 Bornyl acetate 1586 0.44 0.06
35 Terpinen-4-ol 1600 6.97 0.72
36 Myrtenal 1623 0.69 -
37 Allo-aromadendrene 1638 1.06 0.03
38 Trans pinocarveol 1654 0.58 0.50
39 Isoborneol 1668 0.59 -
40 β-Gurjunene 1677 0.66 0.04
41 α-Terpineol 1693 0.53 0.03
42 Borneol 1702 3.67 0.97
43 Germacrene –D 1703 5.03 0.08
44 δ-Cadinene 1755 0.60 0.23
45 γ-Cadinene 1756 6.55 0.67
46 Cuminaldehyde 1773 1.04 3.72
47 Ar-curcumene 1776 1.18 2.49
48 Myrtenol 1791 4.26 -
49 Carveol 1804 1.14 0.05
50 β-bisabolol 2014 0.55 0.09 51 Ledol 2030 2.45 1.03
52 (E)-Nerolidol 2036 0.38 0.02
53 Elemol 2070 0.61 0.05
54 Viridiflorol 2090 0.67 0.09
55 Cadinol 2145 1.14 0.07
56 Spathulenol 2128 5.80 0.11
57 Patchoulene 2154 0.53 0.07
58 τ-Cadinol 2164 3.21 0.70
59 τ-Muurolol 2187 0.19 0.06
60 α-Cadinol 2200 1.98 0.15
61 Carvacrol 2204 2.51 0.11
62 α-Bisabolol 2212 2.36 0.06
63 β-Eudesmol 2233 10.49 0.06
64 Phytol 2603 1.18 0.19
65 NI1 (Molecular weight = 200) 2655 0.20 48.13
66 NI2 (Molecular weight = 200) 2699 0.17 37.31
67 Hexadecanoic acid 2845 0.90 0.88
TOTAL 99.14 99.44
Monoterpenes oxygenated 36.94 2.52
Sesquiterpenes oxygenated 29.83 2.50
Monoterpenes hydrocarbons 7.24 0.22
Sesquiterpenes hydrocarbons 16.83 3.77
Aldehydes 1.82 3.73
Divers 6.10 1.27
NI 0.38 85.44
a RI : retention indices on polar column; (-) : compound absent
Order of elution and percentages (%) of individual components are given on HP Innowax polar column; NI: non identified compounds.
Table 2: Antifungal activity of S. chamaecyparissus flowerhead and root essential oils using percentage inhibition of micro-organisms and minimum inhibitory concentration in µg/ml
Flowerhead essential oil Root essential oil
a b Microorganisms I% MIC I% MIC
Trichophyton rubrum (B) 85.36 500 68 750
Microsporum canis (IP) 73.00 750 30.95 1000
Epidermophyton floccosum (B) 89.25 500 29.03 1000
Aspergillus fumigatus (B) 82.00 500 50.00 1000
Scopulariopsis brevicaulis (B) 85.70 500 42.85 1000
Scytalidium dimidiatum (IP). 83.82 500 44.11 1000
Candida albicans (B) 0.00 1000 0.00 1000
a I%: Percentage inhibition of micro-organisms in the presence of 500 µg/ml of essential oil (0 - 25%, no or little inhibition; 26 - 50%, average inhibition; 51- 75%, strong inhibition); bMIC : Minimum inhibitory concentration (µg/ml); Strains from IP (Institute Pasteur de Paris, France) and from B (Microbiological laboratory; Faculty of Medicine Besancon, France).
Table 3: In vitro antibacterial activity of Santolina chamaecyparissus flowerhead and root essential oils
Growth inhibition zone diameter a MIC c (mm)
Bacteria Flowerhead Root Gentamicin Flowerhead Root Gentamicin oil b oil oil oil
Escherichia coli 15 10 19 1.25 1.25 0.312
(ATCC 25922)
Pseudomonas 7 NA 14 0.625 5 0.312 aeruginosa
(ATCC 27853)
Enterococcus 26 7 23 0.625 2.5 0.312 faecalis
(ATCC 29212)
Staphylococcus 12 12 19.5 2.5 1.25 0.312 aureus
(ATCC 25923)
Citobacter 13 NA 18 10 5 0.625 freundeii
Proteus mirabilis 12 NA 14 10 > 10 1 .25
a Inhibition zones including the diameter of the paper disc (6mm); b 10 µl of essential oil/disc; c MIC: Minimal Inhibition Concentration (µg/ml); NA: no activity. Abstract: The antimicrobial properties of essential oil from various Santolina species have not been enough investigated in the previous studies dealing with the biological activities of medicinal plants. In Tunisia, Santolina chamaecyparissus L. (Asteraceae) is the only Santolina species recorded and is used as vermifuge and emmanagogue. The chemical composition, antibacterial and antifungal properties of essential oils from the flowerheads and roots of spontaneous S. chamaecyparissus growing in Tunisia and the chemical composition which lead to the Tunisian chemotype are investigated here for the first time. Essential oils isolated by hydro distillation from flowerheads and roots of S. chamaecyparissus were analyzed by GC and GC/MS. Two methods served for antimicrobial assays of the essential oils: diffusion in a solid medium and Micro-well dilution assay. Antifungal tests were carried out by the agar incorporation method. Sixty seven constituents were identified from the essential oil of the flowerhead. The major constituents were: 1,8- -eudesmol. Two non identified compounds were present at the highest concentration in root oil. Flowerhead oil was characterized by high contents in monoterpenes and sesquiterpenes oxygenated compounds. The flowerhead essential oil demonstrated potent of antibacterial properties against Pseudomonas aeruginosa ATCC and Enterococcus faecalis ATCC, with MIC of 0.625 µg/ml. These findings demonstrate that the flowerhead essential oils of S. chamaecyparissus have excellent antibacterial properties and for this reason they could contribute to decrease the problem of microbial resistance to antibiotics.
Keywords: antibacterial activity, antifungal activity, Essential oils, secondary metabolites, S. chamaecyparissus L, Asteraceae family